LOCUS       62056_11390        7148 bp DNA     circular SYN 23-MAR-2020
DEFINITION  Cloning vector pGEM-T_blanc_GFPco8g_tGAL_URA3locus_tTK, complete 
            sequence.
ACCESSION   MN271957
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7148)
  AUTHORS   Lodens S, Roelants SLK.W., Geys R, Coussement P, De Maeseneire SL, 
            Soetaert W.
  TITLE     The effect of its gene cluster localization on sophorolipid 
            production by Starmerella bombicola
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7148)
  AUTHORS   Lodens S, Roelants SLK.W.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-AUG-2019) Inbio.be; Department of Biotechnology, Ghent
            University, Coupure links 653, Ghent 9000, Belgium
REFERENCE   3  (bases 1 to 7148)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-AUG-2019) Inbio.be; Department of Biotechnology, Ghent 
            University, Coupure links 653, Ghent 9000, Belgium"
FEATURES             Location/Qualifiers
     source          1..7148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    53..1843
                     /label=5' homologous region ura3 locus
                     /note="5' homologous region ura3 locus"
     regulatory      427..1054
                     /regulatory_class="promoter"
     gene            1055..1843
                     /gene="ura3"
                     /label=ura3
     CDS             1055..1843
                     /codon_start=1
                     /transl_table=11
                     /gene="ura3"
                     /product="URA3"
                     /label=ura3
                     /protein_id="QIM58711.1"
                     /translation="MKLSYEERRERLPAVAARLFDIIVSKQTNLCASLDVRTTSELLSI
                     LDRIGPYICMVKTHIDIIDDFEYDTTVSGLKQLSTKHNFLIFEDRKFADIGSTVKAQYA
                     GGVFKIAQWADITNAHGVPGPGIVSGLEEAAKETTDEPRGLVMLAELSSKGTLAHGEYS
                     QATVDIARSNRAFVFGFIAQQKVGKPEEDWVIMTPGVGLDDKGDGLGQQYRTVDDVIET
                     GTDVIIVGRGLYSKGRDPVHEAQRYQKAGWNAYLRKVQSR"
     polyA_signal    1908..1956
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             1976..2689
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      2693..3161
                     /regulatory_class="terminator"
     misc_feature    3162..4210
                     /label=3' homologous region ura3 locus
                     /note="3' homologous region ura3 locus"
     promoter        complement(4273..4291)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4309..4325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4333..4349)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4357..4387)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4402..4423)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4711..5299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5473..6330)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6331..6435)
                     /label=AmpR promoter
     rep_origin      complement(6513..6968)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7109..7125
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7132..7148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggccgcggga ttagaacaag
       61 gccgagtatg tcgacgtttc cagggacaac agcgacgcta gccatgctga agacagcacc
      121 gctgctatcg agcagaagaa atacgaaatt gattcctaca actcggcctt ggcgcttgag
      181 aagttcccag tagggtggta gcagtccagc cgccaacaga accgcggcga tgactccagg
      241 aattagattg ggccattcaa ttccttttct atataaactg cggagcggga aaatgcaacc
      301 agcctcgacg ccaatgctta cgcatactaa acacgccagt agaatgctgc ttttcgtgcc
      361 agaccagctg acgggcggat agtacaggct ttgccaaaag cctataaggc taaagaaagt
      421 aaacaagtga ggttgaacca tgatggcagt gttcgaattc tgatcaatga agtacactgc
      481 gaagggaatc cccgaaacgg cgaacaaaaa gaacatcaga ggaggaacgc cctcgcaatc
      541 ccgaacatac cagtttcgca gaacctgggg tatcaactgg atgcaccagc atactgttcc
      601 cactgttgcc aatgctgtag acgctccatt gttgtcagtc attttagcat tttacagtaa
      661 ccaactccaa aaaacagccc gctctgctgg gaagacttcg caattattta tccactactg
      721 ctgcggttat atacttctcg atctcagtct cggttataat tgccgcttga cagcctggag
      781 aaattcggat actccacgtg ataattgcca tagggcataa ttttcgaaac agctcgcaac
      841 gatctcggct agttttcccc ttttttgacc catatcgacg ctgagactca ctcacttgat
      901 gcctaccgtt agggtaaatt tttcaagcct gcagaatatc gcgggacgca gtctcctgca
      961 cgcgcgtgac ttcatcttac ttacatcaaa cagcccgatt aatttgaaaa gtcctagctg
     1021 atcgagggca cgggcactac tgtagagaaa taatatgaag ctgagctatg aggagcgccg
     1081 agagaggctg ccggctgtag cagcccggct attcgacatc attgtgagca agcaaacaaa
     1141 tctttgcgca agcttggatg tgcgaactac ctctgagtta ctgagtatcc tggaccgcat
     1201 tggaccttac atttgtatgg ttaagaccca cattgacata attgacgact tcgaatacga
     1261 cacaactgtc agcggtttga aacagctttc aacgaagcac aattttctca tttttgaaga
     1321 ccgaaagttc gcagacatcg gttccactgt taaggcccaa tatgcaggtg gagtgtttaa
     1381 gatcgctcaa tgggctgata taacaaatgc tcacggtgtt cctgggccgg gaattgtgag
     1441 cggactagaa gaggctgcga aggaaactac ggatgaacct cgcggccttg tcatgcttgc
     1501 agaactgagt tcgaagggca cactggctca cggcgaatac tcgcaagcga cagtagacat
     1561 cgctcgcagt aaccgcgcat ttgtgtttgg tttcatcgct cagcaaaaag tcggaaagcc
     1621 agaggaagac tgggtcatta tgactcctgg ggtgggcctg gacgacaaag gtgatggatt
     1681 ggggcagcag tatcgtactg tggacgacgt catagagacc ggcacagacg ttattatcgt
     1741 cggacgcggg ctctatagca agggacgaga tcctgtgcac gaagctcagc gttaccaaaa
     1801 ggcgggctgg aatgcatatc tgagaaaagt tcagtcaaga tgagggagat gggggaggct
     1861 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     1921 acagaataaa acgcacgggt gttgggtcgt ttgttcaaat cactagtata cccggatgtc
     1981 gaaaggagag gagctgttca ctggcgtcgt cccaatcctg gtcgaacttg atggcgacgt
     2041 taacggtcac aagtttagcg tgagcggtga aggtgagggt gatgctacct acggcaagct
     2101 gactcttaag ttcatctgca ccactggcaa gcttccagtt ccatggccca cactcgttac
     2161 aaccttcggc tatggcgttc agtgctttgc ccgttacccc gaccacatga agcagcacga
     2221 cttcttcaag tcggccatgc ctgagggcta tgttcaggaa cgtactatct tcttcaagga
     2281 cgacggcaac tacaaaactc gtgctgaagt caagtttgag ggcgatactt tggtaaaccg
     2341 tatcgagctc aagggcattg acttcaagga ggacggtaac attcttggtc ataagcttga
     2401 gtataattac aactctcaca acgtctacat tatggctgac aagcagaaga atggaattaa
     2461 agtcaacttc aagattcgcc ataacatcga ggacggcagc gtacagctcg ctgaccacta
     2521 ccaacagaac actcccattg gcgacggccc tgttcttctc cctgacaacc actatctatc
     2581 tacccagtcc gccttgtcca aagaccccaa cgagaagcgc gaccacatgg ttcttttgga
     2641 gttcgttact gccgctggca tcacgcacgg aatggacgag ctgtacaaat aactgcagtt
     2701 ttaaaaattt gcagctctat agtaatttag ttttgtatgg tttatagaaa cttagtttaa
     2761 aaaataagat aggttatgtt tttacgacac agtcgtccta gaatcagatc aacatcctgt
     2821 aagtggcacg ttgagtagaa gtgattattc atgggatatt aagaaaacaa aaattgagct
     2881 cagaggaatt cgattaaata tacactagat gctatcgata atatcgttaa accagaaccc
     2941 agagagaaca acaacataga tcaatatcaa aatgacacct ttaaagtagt ttgatttacc
     3001 ctcggcgtag atgtaggtga acaagtatat ggaaatgagt gtggcgatga tatcccacag
     3061 tgggaaaatc aacgtgaatg tctgctcgac attttcaaag ttctggtata ttgagtagag
     3121 gaccaagcac ggaatttgga tcaacaccac ctgcaaagcg tatggatccc cgcagggcat
     3181 gcaacttgca catgaatacc tataaaatct gattaaatta ccataaaagg tacagattaa
     3241 aatatatatg ccttcaatgg catccttcgc gattctgatt cgtcagcaca cttcaacctt
     3301 cctactatga gtgacagtga tgatgatctg ctggcattgg ccgacgttgg ctccgactcc
     3361 gaagaggaaa tctcgcagcc gtcgccgcca agcaatgagg tcgtcaatcc ctatcctcta
     3421 gaaggcaaat atctcgatgc tgaagacagg gcgaagttgg acgcgctgcc agagattgag
     3481 cgagaagaga tcttgtatga ccgagctcag gagatgcagc ggtacgagga gagaaggtat
     3541 cttgctcagc gaaggaagca gatgacgcgg gttgctgacg aggacgaagc cccctccgcc
     3601 aagcgtcaac ggggtacaac aggcgtctct tcgggtacga agtcatctct tgaggcatta
     3661 aagaaacgaa gggcccagca gtctcggaag tcctcacgcc atggagttga tgacgatgtg
     3721 tatagtgacg atgatgaagg caatgacagt tcctcggcat cggaaggctc agatattgac
     3781 tcggaagatg aagaggtaag gcgaagtttc cgtcggcgaa agacagacga agttgagtgg
     3841 gctgataatg atgacgacct tgcggaggag gttatctccc tgagcgatat caataaaata
     3901 cgttggggca aatcgctgtt tgcacggttc agcatgatgc ccggcttcgt tgatgcagtg
     3961 cctggctgct ttgtgagagt taatattgga gctaaccgct ccggacagcc tgtttatcgg
     4021 ctctgtcaga tcaaaaaggt cgtgacccat cgcccgtacc agtttatgga taagatggtt
     4081 gacgaggcca ttgttgcgtc catggctggc tcagaaagaa ctcttgaatt cagtatatgc
     4141 tccgactcgc ctgcttctca agccgaatac gatttttgga aaacggctat gtcggaggct
     4201 aatctgacgc ggccgcctgc aggtcgacca tatgggagag ctcccaacgc gttggatgca
     4261 tagcttgagt attctatagt gtcacctaaa tagcttggcg taatcatggt catagctgtt
     4321 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4381 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4441 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4501 ggggagaggc ggtttgcgta ttgggcgctc ctccgcttcc tcgctcactg actcgctgcg
     4561 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4621 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4681 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4741 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4801 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4861 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4921 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4981 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5041 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5101 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5161 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5221 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5281 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5341 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5401 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5461 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5521 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5581 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5641 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5701 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5761 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5821 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5881 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5941 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6001 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6061 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6121 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6181 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6241 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6301 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6361 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6421 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gatgcggtgt gaaataccgc
     6481 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     6541 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6601 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6661 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6721 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6781 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6841 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6901 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6961 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     7021 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     7081 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
     7141 tcactata
//
